LOCUS       sequence-324610-       12589 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Coexpression plasmid of QuasAr2 and CheRiff for all-optical 
            electrophysiology.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12589)
  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 12589)
  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: DRH296: FCK-Optopatch2
FEATURES             Location/Qualifiers
     source          1..12589
                     /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"
     primer_bind     772..792
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     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"
     promoter        2602..3890
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     CDS             3903..4661
                     /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             4668..5375
                     /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             5469..5525
                     /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             5526..6470
                     /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(6540..6542,6543..7256)
                     /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(6540..6542,6543..7253)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(6582..6603)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6843..6862)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7190..7211
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    7281..7869
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7334..7354)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7752..7763)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(7872..7888)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7873..7889)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             8394..8574
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(8600..8617)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8606..8830
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      8876..9304
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8963..8982)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9173..9194
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9313..9333)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        9318..9647
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      9498..9633
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     9560..9579
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        9695..9742
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             9761..10135
                     /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    10265..10386
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10302..10321)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10356..10375
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(10435..10451)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10435..10451)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10448..10470)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10459..10475
                     /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(10483..10489,10490..10507,10508..10513))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10528..10549
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10666..10683)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10837..11425)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10917..10936)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11596..12387,12388..12456))
                     /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     12219..12238
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12457..12561)
                     /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 acattatggc cttaggtcac ttcatctcca tggggttctt
     2641 cttctgattt tctagaaaat gagatggggg tgcagagagc ttcctcagtg acctgcccag
     2701 ggtcacatca gaaatgtcag agctagaact tgaactcaga ttactaatct taaattccat
     2761 gccttggggg catgcaagta cgatatacag aaggagtgaa ctcattaggg cagatgacca
     2821 atgagtttag gaaagaagag tccagggcag ggtacatcta caccacccgc ccagccctgg
     2881 gtgagtccag ccacgttcac ctcattatag ttgcctctct ccagtcctac cttgacggga
     2941 agcacaagca gaaactggga caggagcccc aggagaccaa atcttcatgg tccctctggg
     3001 aggatgggtg gggagagctg tggcagaggc ctcaggaggg gccctgctgc tcagtggtga
     3061 cagatagggg tgagaaagca gacagagtca ttccgtcagc attctgggtc tgtttggtac
     3121 ttcttctcac gctaaggtgg cggtgtgata tgcacaatgg ctaaaaagca gggagagctg
     3181 gaaagaaaca aggacagaga cagaggccaa gtcaaccaga ccaattccca gaggaagcaa
     3241 agaaaccatt acagagacta caagggggaa gggaaggaga gatgaattag cttcccctgt
     3301 aaaccttaga acccagctgt tgccagggca acggggcaat acctgtctct tcagaggaga
     3361 tgaagttgcc agggtaacta catcctgtct ttctcaagga ccatcccaga atgtggcacc
     3421 cactagccgt taccatagca actgcctctt tgccccactt aatcccatcc cgtctgttaa
     3481 aagggcccta tagttggagg tgggggaggt aggaagagcg atgatcactt gtggactaag
     3541 tttgttcgca tccccttctc caaccccctc agtacatcac cctgggggaa cagggtccac
     3601 ttgctcctgg gcccacacag tcctgcagta ttgtgtatat aaggccaggg caaagaggag
     3661 caggttttaa agtgaaaggc aggcaggtgt tggggaggca gttaccgggg caacgggaac
     3721 agggcgtttc ggaggtggtt gccatgggga cctggatgct gacgaaggct cgcgaggctg
     3781 tgagcagcca cagtgccctg ctcagaagcc ccaagctcgt cagtcaagcc ggttctccgt
     3841 ttgcactcag gagcacgggc aggcgagtgg cccctagttc tgggggcagc gggggatcca
     3901 ccatggtaag tatcgctctg caggctggtt acgacctact gggtgacggc agacctgaaa
     3961 ccctgtggct gggcatcggc actctgctga tgctgattgg aaccttctac tttctggtcc
     4021 gcggatgggg agtcaccgat aaggatgccc gggaatatta cgctgtgact atcctggtgt
     4081 ccggaatcgc atccgccgca tatctgtcta tgttctttgg tatcgggctt actgaggtgt
     4141 ccgtcggggg cgaaatgttg gatatctatt atgccaggta cgcccagtgg ctgtttacca
     4201 ccccacttct gctgctgcac ctggcccttc tcgctaaggt ggatcgggtg accatcggca
     4261 ccctggtggg tgtggacgcc ctgatgatcg tcactggcct catcggagcc ttgagccaca
     4321 cggccatagc cagatacagt tggtggttgt tctctacaat ttgcatgata gtggtgctct
     4381 atgttctggc tacatccctg cgatctgctg caaaggagcg gggccccgag gtggcatcta
     4441 cctttaacac cctgacagct ctggtcttgg tgctgtggac cgcttaccct atcctgtgga
     4501 tcataggcac tgagggcgct ggcgtggtgg gcctgggcat cgaaactctg ctgtttatgg
     4561 tgttggacgt gactgccaag gtcggctttg gctttatcct gttgagatcc cgggctattc
     4621 tgggcgacac cgaggcacca gaacccagtg ccggtgccga tacgttgatg gtgagcaagg
     4681 gcgaggagta taacatggcc atcatcaagg agttcatgcg cttcaaggtg cgcatggagg
     4741 gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg
     4801 gctttcagac cgctaagctg aaggtgacca agggtggccc cctgcccttc gcctgggaca
     4861 tcctgtcccc tcatttcacc tacggctcca aggcctacgt gaagcacccc gccgacatcc
     4921 ccgactactt caagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaactacg
     4981 aggacggcgg cgtggtgacc gtgacccagg actcctccct gcaggacggc gagttcatct
     5041 acaaggtgaa gctgcgcggc accaacttcc cctccgacgg ccccgtgatg cagaagaaga
     5101 ccatgggctg ggaggcctcc tccgagcgga tgtaccccga ggacggtgcc ctgaagggca
     5161 agatcaagat gaggctgaag ctgaaggacg gcggccacta cacctccgag gtcaagacca
     5221 cctacaaggc caagaagccc gtgcagctgc ccggcgccta catcgtcgac atcaagttgg
     5281 acatcacctc ccacaacgag gactacacca tcgtggaaca gtacgaacgc gccgagggcc
     5341 gccactccac cggcggcatg gacgagctgt acaagagtag aatcacaagc gaaggcgagt
     5401 acatccccct ggatcaaata gacataaatg taggtggatt ttgttatgag aatgaagtgg
     5461 gctccggagc cacgaacttc tctctgttaa agcaagcagg agacgtggaa gaaaaccccg
     5521 gtcccggcgg agctcctgct ccagacgctc acagcgcccc acctggaaac gattctgccg
     5581 gaggcagtga gtaccatgcc ccagctggat atcaagtgaa tccaccctac caccccgtgc
     5641 atgggtatga ggaacagtgc agctccatct acatctacta tggggccctg tgggagcagg
     5701 aaacagctag gggcttccag tggtttgccg tgttcctgtc tgccctgttt ctggctttct
     5761 acggctggca cgcctataag gccagcgtgg gatgggagga agtgtacgtg tgctccgtgg
     5821 agctgatcaa agtgattctg gagatctatt tcgagttcac cagtcctgct atgctgttcc
     5881 tgtacggagg gaacattacc ccatggctga gatatgccga atggctgctg acatgtcccg
     5941 tgatcctgat tcatctgtct aacatcaccg gcctgagtga ggcatacaat aagcggacaa
     6001 tggctctgct ggtgtccgac ctgggaacta tttgcatggg agtgacagcc gctctggcca
     6061 ctgggtgggt gaagtggctg ttttactgta tcggcctggt gtatggaacc cagacattct
     6121 acaacgctgg aatcatctac gtggagtctt actatatcat gcctgccggc ggctgtaaga
     6181 aactggtgct ggccatgact gccgtgtact attctagttg gctgatgttt cccggcctgt
     6241 tcatctttgg gcctgaaggc atgcacaccc tgagcgtggc tgggtccact attggccata
     6301 ccatcgccga cctgctgtcc aagaatattt ggggactgct ggggcacttc ctgcggatca
     6361 aaattcacga gcatatcatt atgtacggcg atatcaggag accagtgagc tcccagtttc
     6421 tgggacgcaa ggtggacgtg ctggccttcg tgacagagga agataaagtg gcggccgcca
     6481 agagcaggat caccagcgag ggcgagtaca tccccctgga ccagatcgac atcaacgtgg
     6541 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
     6601 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
     6661 agctgaccct gaagttcatt tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     6721 tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc
     6781 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
     6841 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     6901 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     6961 tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca
     7021 tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc
     7081 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     7141 tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     7201 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaagaat
     7261 tcgatatcaa gcttatcgat aatcaacctc tggattacaa aatttgtgaa agattgactg
     7321 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt
     7381 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc
     7441 tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt
     7501 ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga
     7561 ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct
     7621 gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat
     7681 cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct
     7741 gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc
     7801 tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg
     7861 cctccccgca tcgataccgt cgacctcgag acctagaaaa acatggagca atcacaagta
     7921 gcaatacagc agctaccaat gctgattgtg cctggctaga agcacaagag gaggaggagg
     7981 tgggttttcc agtcacacct caggtacctt taagaccaat gacttacaag gcagctgtag
     8041 atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa
     8101 gacaagatat ccttgatctg tggatctacc acacacaagg ctacttccct gattggcaga
     8161 actacacacc agggccaggg atcagatatc cactgacctt tggatggtgc tacaagctag
     8221 taccagttga gcaagagaag gtagaagaag ccaatgaagg agagaacacc cgcttgttac
     8281 accctgtgag cctgcatggg atggatgacc cggagagaga agtattagag tggaggtttg
     8341 acagccgcct agcatttcat cacatggccc gagagctgca tccggactgt actgggtctc
     8401 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
     8461 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
     8521 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagggccc
     8581 gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     8641 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     8701 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     8761 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     8821 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg
     8881 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     8941 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     9001 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     9061 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     9121 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     9181 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     9241 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     9301 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     9361 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     9421 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     9481 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     9541 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     9601 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     9661 cttgtatatc cattttcgga tctgatcagc acgtgttgac aattaatcat cggcatagta
     9721 tatcggcata gtataatacg acaaggtgag gaactaaacc atggccaagt tgaccagtgc
     9781 cgttccggtg ctcaccgcgc gcgacgtcgc cggagcggtc gagttctgga ccgaccggct
     9841 cgggttctcc cgggacttcg tggaggacga cttcgccggt gtggtccggg acgacgtgac
     9901 cctgttcatc agcgcggtcc aggaccaggt ggtgccggac aacaccctgg cctgggtgtg
     9961 ggtgcgcggc ctggacgagc tgtacgccga gtggtcggag gtcgtgtcca cgaacttccg
    10021 ggacgcctcc gggccggcca tgaccgagat cggcgagcag ccgtgggggc gggagttcgc
    10081 cctgcgcgac ccggccggca actgcgtgca cttcgtggcc gaggagcagg actgacacgt
    10141 gctacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
    10201 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
    10261 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
    10321 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
    10381 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
    10441 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
    10501 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
    10561 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
    10621 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
    10681 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
    10741 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
    10801 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
    10861 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
    10921 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
    10981 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
    11041 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
    11101 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
    11161 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
    11221 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
    11281 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
    11341 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
    11401 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
    11461 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
    11521 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
    11581 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
    11641 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
    11701 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
    11761 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
    11821 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
    11881 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
    11941 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
    12001 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
    12061 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
    12121 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
    12181 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
    12241 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
    12301 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
    12361 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
    12421 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
    12481 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
    12541 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgac
//
