LOCUS       sequence-104116-       11895 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Coexpression plasmid of QuasAr2 and CheRiff for all-optical 
            electrophysiology under CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11895)
  AUTHORS   Hochbaum DR, Zhao Y, Farhi SL, Klapoetke N, Werley CA, Kapoor V, Zou
            P, Kralj JM, Maclaurin D, Smedemark-Margulies N, Saulnier JL, 
            Boulting GL, Straub C, Cho YK, Melkonian M, Wong GK, Harrison DJ, 
            Murthy VN, Sabatini BL, Boyden ES, Campbell RE, Cohen AE
  TITLE     All-optical electrophysiology in mammalian neurons using engineered 
            microbial rhodopsins.
  JOURNAL   Nat Methods. 2014 Aug;11(8):825-33. doi: 10.1038/nmeth.3000. Epub 
            2014 Jun 22.
  PUBMED    24952910
REFERENCE   2  (bases 1 to 11895)
  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"; doi: 
            "10.1038/nmeth.3000"; journalName: "Nat Methods"; date: "2014-08"; 
            volume: "11"; issue: "8"; pages: "825-33"
COMMENT     Sequence Label: pMOS001: CMV-Optopatch2_FCK
FEATURES             Location/Qualifiers
     source          1..11895
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             835..1015
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1680..1913
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2098..2142
                     /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    2440..2557
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        2616..2919
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2920..3123
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3120..3144
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3209..3967
                     /codon_start=1
                     /product="achaerhodopsin-3 (Arch) derivative with improved 
                     voltage sensitivity (Hochbaum et al., 2014)"
                     /label=QuasAr2
                     /translation="MVSIALQAGYDLLGDGRPETLWLGIGTLLMLIGTFYFLVRGWGVT
                     DKDAREYYAVTILVSGIASAAYLSMFFGIGLTEVSVGGEMLDIYYARYAQWLFTTPLLL
                     LHLALLAKVDRVTIGTLVGVDALMIVTGLIGALSHTAIARYSWWLFSTICMIVVLYVLA
                     TSLRSAAKERGPEVASTFNTLTALVLVLWTAYPILWIIGTEGAGVVGLGIETLLFMVLD
                     VTAKVGFGFILLRSRAILGDTEAPEPSAGAD"
     CDS             3974..4681
                     /codon_start=1
                     /product="photostable monomeric orange derivative of DsRed 
                     fluorescent protein (Shaner et al., 2008)"
                     /label=mOrange2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEYNMAIIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEG
                     FQTAKLKVTKGGPLPFAWDILSPHFTYGSKAYVKHPADIPDYFKLSFPEGFKWERVMNY
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GKIKMRLKLKDGGHYTSEVKTTYKAKKPVQLPGAYIVDIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     CDS             4775..4831
                     /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             4832..5776
                     /codon_start=1
                     /product="blue light-activated optogenetic actuator derived
                     from Scherffelia dubia channelrhodopsin (Hochbaum et al., 
                     2014)"
                     /label=CheRiff
                     /translation="GGAPAPDAHSAPPGNDSAGGSEYHAPAGYQVNPPYHPVHGYEEQC
                     SSIYIYYGALWEQETARGFQWFAVFLSALFLAFYGWHAYKASVGWEEVYVCSVELIKVI
                     LEIYFEFTSPAMLFLYGGNITPWLRYAEWLLTCPVILIHLSNITGLSEAYNKRTMALLV
                     SDLGTICMGVTAALATGWVKWLFYCIGLVYGTQTFYNAGIIYVESYYIMPAGGCKKLVL
                     AMTAVYYSSWLMFPGLFIFGPEGMHTLSVAGSTIGHTIADLLSKNIWGLLGHFLRIKIH
                     EHIIMYGDIRRPVSSQFLGRKVDVLAFVTEEDKV"
     CDS             join(5846..5848,5849..6562)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(5846..5848,5849..6559)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(5888..5909)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6149..6168)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6496..6517
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    6587..7175
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6640..6660)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7058..7069)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(7178..7194)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7179..7195)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             7700..7880
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(7906..7923)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7912..8136
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      8182..8610
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8269..8288)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8479..8500
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8619..8639)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        8624..8953
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8804..8939
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     8866..8885
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        9001..9048
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             9067..9441
                     /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    9571..9692
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9608..9627)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9662..9681
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(9741..9757)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9741..9757)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9754..9776)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9765..9781
                     /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(9789..9795,9796..9813,9814..9819))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9834..9855
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9972..9989)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10143..10731)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10223..10242)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10902..11693,11694..11762))
                     /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     11525..11544
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11763..11867)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
      841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     1501 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg
     1561 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     1621 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     1681 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     1741 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     1801 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     1861 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     1921 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     1981 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     2041 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     2101 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
     2161 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
     2221 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
     2281 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa
     2341 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc ggcactgcgt
     2401 gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat
     2461 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa
     2521 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag
     2581 agatccagtt tggttaatta agcttgggag ttccgcgtta cataacttac ggtaaatggc
     2641 ccgcctggct gaccgcccaa cgacccccgc ccattgacga caataatgac gtatgttccc
     2701 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
     2761 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
     2821 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
     2881 tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac
     2941 atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac
     3001 gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac
     3061 tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga
     3121 gctcgtttag tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat
     3181 agaagacacc gactctagag gatccaccat ggtaagtatc gctctgcagg ctggttacga
     3241 cctactgggt gacggcagac ctgaaaccct gtggctgggc atcggcactc tgctgatgct
     3301 gattggaacc ttctactttc tggtccgcgg atggggagtc accgataagg atgcccggga
     3361 atattacgct gtgactatcc tggtgtccgg aatcgcatcc gccgcatatc tgtctatgtt
     3421 ctttggtatc gggcttactg aggtgtccgt cgggggcgaa atgttggata tctattatgc
     3481 caggtacgcc cagtggctgt ttaccacccc acttctgctg ctgcacctgg cccttctcgc
     3541 taaggtggat cgggtgacca tcggcaccct ggtgggtgtg gacgccctga tgatcgtcac
     3601 tggcctcatc ggagccttga gccacacggc catagccaga tacagttggt ggttgttctc
     3661 tacaatttgc atgatagtgg tgctctatgt tctggctaca tccctgcgat ctgctgcaaa
     3721 ggagcggggc cccgaggtgg catctacctt taacaccctg acagctctgg tcttggtgct
     3781 gtggaccgct taccctatcc tgtggatcat aggcactgag ggcgctggcg tggtgggcct
     3841 gggcatcgaa actctgctgt ttatggtgtt ggacgtgact gccaaggtcg gctttggctt
     3901 tatcctgttg agatcccggg ctattctggg cgacaccgag gcaccagaac ccagtgccgg
     3961 tgccgatacg ttgatggtga gcaagggcga ggagtataac atggccatca tcaaggagtt
     4021 catgcgcttc aaggtgcgca tggagggctc cgtgaacggc cacgagttcg agatcgaggg
     4081 cgagggcgag ggccgcccct acgagggctt tcagaccgct aagctgaagg tgaccaaggg
     4141 tggccccctg cccttcgcct gggacatcct gtcccctcat ttcacctacg gctccaaggc
     4201 ctacgtgaag caccccgccg acatccccga ctacttcaag ctgtccttcc ccgagggctt
     4261 caagtgggag cgcgtgatga actacgagga cggcggcgtg gtgaccgtga cccaggactc
     4321 ctccctgcag gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc
     4381 cgacggcccc gtgatgcaga agaagaccat gggctgggag gcctcctccg agcggatgta
     4441 ccccgaggac ggtgccctga agggcaagat caagatgagg ctgaagctga aggacggcgg
     4501 ccactacacc tccgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg
     4561 cgcctacatc gtcgacatca agttggacat cacctcccac aacgaggact acaccatcgt
     4621 ggaacagtac gaacgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa
     4681 gagtagaatc acaagcgaag gcgagtacat ccccctggat caaatagaca taaatgtagg
     4741 tggattttgt tatgagaatg aagtgggctc cggagccacg aacttctctc tgttaaagca
     4801 agcaggagac gtggaagaaa accccggtcc cggcggagct cctgctccag acgctcacag
     4861 cgccccacct ggaaacgatt ctgccggagg cagtgagtac catgccccag ctggatatca
     4921 agtgaatcca ccctaccacc ccgtgcatgg gtatgaggaa cagtgcagct ccatctacat
     4981 ctactatggg gccctgtggg agcaggaaac agctaggggc ttccagtggt ttgccgtgtt
     5041 cctgtctgcc ctgtttctgg ctttctacgg ctggcacgcc tataaggcca gcgtgggatg
     5101 ggaggaagtg tacgtgtgct ccgtggagct gatcaaagtg attctggaga tctatttcga
     5161 gttcaccagt cctgctatgc tgttcctgta cggagggaac attaccccat ggctgagata
     5221 tgccgaatgg ctgctgacat gtcccgtgat cctgattcat ctgtctaaca tcaccggcct
     5281 gagtgaggca tacaataagc ggacaatggc tctgctggtg tccgacctgg gaactatttg
     5341 catgggagtg acagccgctc tggccactgg gtgggtgaag tggctgtttt actgtatcgg
     5401 cctggtgtat ggaacccaga cattctacaa cgctggaatc atctacgtgg agtcttacta
     5461 tatcatgcct gccggcggct gtaagaaact ggtgctggcc atgactgccg tgtactattc
     5521 tagttggctg atgtttcccg gcctgttcat ctttgggcct gaaggcatgc acaccctgag
     5581 cgtggctggg tccactattg gccataccat cgccgacctg ctgtccaaga atatttgggg
     5641 actgctgggg cacttcctgc ggatcaaaat tcacgagcat atcattatgt acggcgatat
     5701 caggagacca gtgagctccc agtttctggg acgcaaggtg gacgtgctgg ccttcgtgac
     5761 agaggaagat aaagtggcgg ccgccaagag caggatcacc agcgagggcg agtacatccc
     5821 cctggaccag atcgacatca acgtggtgag caagggcgag gagctgttca ccggggtggt
     5881 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
     5941 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
     6001 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag
     6061 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     6121 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
     6181 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     6241 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
     6301 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga
     6361 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     6421 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa
     6481 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
     6541 catggacgag ctgtacaagt aagaattcga tatcaagctt atcgataatc aacctctgga
     6601 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     6661 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     6721 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     6781 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     6841 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     6901 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     6961 ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac
     7021 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     7081 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     7141 gacgagtcgg atctcccttt gggccgcctc cccgcatcga taccgtcgac ctcgagacct
     7201 agaaaaacat ggagcaatca caagtagcaa tacagcagct accaatgctg attgtgcctg
     7261 gctagaagca caagaggagg aggaggtggg ttttccagtc acacctcagg tacctttaag
     7321 accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact
     7381 ggaagggcta attcactccc aacgaagaca agatatcctt gatctgtgga tctaccacac
     7441 acaaggctac ttccctgatt ggcagaacta cacaccaggg ccagggatca gatatccact
     7501 gacctttgga tggtgctaca agctagtacc agttgagcaa gagaaggtag aagaagccaa
     7561 tgaaggagag aacacccgct tgttacaccc tgtgagcctg catgggatgg atgacccgga
     7621 gagagaagta ttagagtgga ggtttgacag ccgcctagca tttcatcaca tggcccgaga
     7681 gctgcatccg gactgtactg ggtctctctg gttagaccag atctgagcct gggagctctc
     7741 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt
     7801 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc
     7861 agtgtggaaa atctctagca gggcccgttt aaacccgctg atcagcctcg actgtgcctt
     7921 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     7981 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     8041 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     8101 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct
     8161 ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     8221 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     8281 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     8341 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     8401 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     8461 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     8521 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     8581 agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg
     8641 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     8701 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     8761 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     8821 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     8881 cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     8941 aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcagcacgt
     9001 gttgacaatt aatcatcggc atagtatatc ggcatagtat aatacgacaa ggtgaggaac
     9061 taaaccatgg ccaagttgac cagtgccgtt ccggtgctca ccgcgcgcga cgtcgccgga
     9121 gcggtcgagt tctggaccga ccggctcggg ttctcccggg acttcgtgga ggacgacttc
     9181 gccggtgtgg tccgggacga cgtgaccctg ttcatcagcg cggtccagga ccaggtggtg
     9241 ccggacaaca ccctggcctg ggtgtgggtg cgcggcctgg acgagctgta cgccgagtgg
     9301 tcggaggtcg tgtccacgaa cttccgggac gcctccgggc cggccatgac cgagatcggc
     9361 gagcagccgt gggggcggga gttcgccctg cgcgacccgg ccggcaactg cgtgcacttc
     9421 gtggccgagg agcaggactg acacgtgcta cgagatttcg attccaccgc cgccttctat
     9481 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     9541 gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac
     9601 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     9661 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc
     9721 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     9781 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     9841 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     9901 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     9961 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
    10021 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
    10081 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
    10141 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
    10201 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
    10261 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
    10321 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
    10381 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
    10441 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
    10501 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
    10561 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
    10621 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
    10681 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
    10741 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
    10801 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
    10861 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
    10921 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
    10981 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
    11041 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
    11101 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
    11161 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
    11221 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
    11281 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
    11341 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
    11401 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
    11461 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
    11521 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
    11581 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
    11641 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
    11701 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
    11761 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
    11821 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
    11881 aaagtgccac ctgac
//
